JP7464591B2 - Metabolite of [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine - Google Patents

Description

The present invention relates to metabolites of [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine (also known as azeliragon). These metabolites can act as RAGE antagonists. These metabolites can also be useful in assays to measure the presence or amount of the metabolite or parent compound in a sample.

The receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin superfamily of cell surface molecules. Activation of RAGE in various tissues and organs has several pathophysiological consequences. RAGE has been implicated in a variety of conditions, including acute and chronic inflammation (Hofmann et. al., Cell 97:889-901 (1999)), the development of late complications of diabetes, such as vascular hyperpermeability (Wautier et. al., J. Clin. Invest. 97:238-243 (1995)), nephropathy (Teillet et. al., J. Am. Soc. Nephrol. 11:1488-1497 (2000)), atherosclerosis (Vlassara et. al., The Finnish Medical Society DUODECIM, Ann. Med. 28:419-426 (1996)), and retinopathy (Hammes et. al., Diabetologia 42:603-607 (1999)). RAGE has also been implicated in Alzheimer's disease (Yan et. al., Nature 382:685-691 (1996)), erectile dysfunction, and tumor invasion and metastasis (Taguchi et. al., Nature 405:354-357 (2000)).

Binding of ligands such as advanced glycation end products (AGEs), S100/calgranulin/EN-RAGE, β-amyloid, CML (N ^ε -carboxymethyllysine), and amphoterin to RAGE has been shown to alter the expression of various genes. For example, in many cell types, the interaction between RAGE and its ligands generates oxidative stress, which leads to the activation of the free radical-sensitive transcription factor NF-κB and the activation of NF-κB-regulated genes such as the cytokines IL-1β, TNF-α, etc. In addition to this, several other regulatory pathways are also activated, such as those involving p21ras.

The MAP kinases ERK1 and ERK2 have been shown to be activated by the binding of AGEs and other ligands to RAGE. Indeed, the transcription of RAGE itself is regulated, at least in part, by NF-κB. Thus, an upward, often detrimental spiral is stimulated by a positive feedback loop initiated by ligand binding. Thus, antagonizing the binding of physiological ligands to RAGE is the target in order to downregulate the pathophysiological changes brought about by excessive concentrations of AGEs and other ligands of RAGE.

Certain RAGE antagonists, including [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine (Compound I), shown below, are disclosed in U.S. Pat. No. 7,361,678.

New agents that antagonize RAGE are continually needed to develop new, more effective pharmaceuticals for treating a variety of diseases. The metabolites, compositions, and methods described herein are directed to these and other objectives.

Hofmann et al., Cell 97:889-901 (1999) Wautier et al. al. , J. Clin. Invest. 97:238-243 (1995) Teillet et al., J. Am. Soc. Nephrol. 11:1488-1497 (2000) Vlassara et al., The Finnish Medical Society DUODECIM, Ann. Med. 28:419-426 (1996) Hammes et al., Diabetologia 42:603-607 (1999) Yan et al., Nature 382:685-691 (1996) Taguchi et al., Nature 405:354-357 (2000)

The present invention provides a compound selected from the group consisting of:
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine;
4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol;
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol;
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol; and 3-(4-{2-{butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol;
or a pharma- ceutically acceptable salt thereof.

The present invention also provides methods for making the compounds of the invention, or a pharma- ceutically acceptable salt thereof.
The present invention further provides pharmaceutical compositions comprising a compound of the invention, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, and methods for making the same.

The present invention also provides methods of using the compounds of the present invention, or pharma- ceutically acceptable salts thereof, as antagonists of RAGE or in the treatment of various medical conditions, including, but not limited to, chronic inflammation, amyloidosis, Alzheimer's disease, cancer, tumor invasion and metastasis, renal failure, or autoimmune-associated inflammation, inflammatory bowel disease, rheumatoid arthritis, psoriasis, multiple sclerosis, hypoxia, stroke, heart attack, hemorrhagic shock, sepsis, organ transplantation, the development of diabetic complications, such as vascular hyperpermeability, diabetic nephropathy, diabetic retinopathy, diabetic foot ulcers, cardiovascular complications, diabetic neuropathy, wound healing disorders, erectile dysfunction, and osteoporosis.

The invention also provides methods of using the compounds of the invention or pharma- ceutically acceptable salts thereof in in vitro or in vivo assays. The in vitro or in vivo assays can be useful for determining (either directly or indirectly) pharmacokinetic parameters of Compound I or the compounds of the invention, e.g., determining the rate of absorption, determining the amount in a tissue or plasma sample, determining the rate of metabolism, or determining the rate of excretion of Compound I or the compounds of the invention.

These and other aspects of the invention are described in more detail in the detailed description of the invention below.

The present invention provides compounds that are metabolites of Compound I, which can bind to RAGE and have activity as RAGE antagonists. Compounds that modulate the activity of RAGE can be useful, for example, in treating a variety of diseases associated with RAGE signaling and/or inflammatory cascades. Compounds of the present invention can also be useful in in vitro or in vivo assays for measuring the pharmacokinetics of Compound I or compounds of the present invention. Compounds of the present invention are listed in Table 1 below.

Metabolites M1, M2, M3, M5, M6 and M7 can be isolated from human plasma and/or bile taken from subjects administered one or more doses of Compound I.
The binding affinities (in the form of dissociation constants Kd) of metabolites M1, M2, M3, M5, M6 and M7 to sRAGE (the soluble extracellular portion of the receptor) are shown below in Table 2. The binding affinity of a compound to sRAGE can predict the activity and potency of the compound as a RAGE antagonist.

The present invention also includes pharma- ceutically acceptable salts of the compounds described herein. As used herein, "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound has been modified by converting an acid or base moiety present therein to its salt form. Examples of pharma- ceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharma- ceutically acceptable salts of the present invention include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharma- ceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from the parent compound containing a basic or acidic moiety. In general, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

The phrase "pharmacologically acceptable" is employed herein to refer to compounds, materials, compositions, and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, consistent with a reasonable benefit/risk ratio.

The compounds of the invention also include all isotopes of atoms present in the metabolites. Isotopes include atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

Synthesis The compounds of the invention, including their salts, can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes. Exemplary synthetic methods for preparing the compounds of the invention are provided in the Examples section below.

Methods of Use As used herein, the term "RAGE mediated disease" is used to refer to one or more conditions, diseases or disease states including, but not limited to, inflammation of the skin, e.g., psoriasis, rheumatoid arthritis, atopic dermatitis, and inflammation of the lungs, e.g., acute or chronic inflammation including asthma and chronic obstructive pulmonary disease, diabetes, diabetes related complications, renal failure, hyperlipidemic atherosclerosis associated with diabetes, neuronal toxicity, restenosis, Down's syndrome, dementia associated with head trauma, amyotrophic lateral sclerosis, multiple sclerosis, amyloidosis, autoimmune diseases, e.g., inflammation associated with autoimmunity or organ, tissue or cell transplantation, wound healing disorders, periodontal disease, neuropathy, neuronal degeneration, vascular permeability, nephropathy, atherosclerosis, retinopathy, Alzheimer's disease, erectile dysfunction, tumor invasion and/or metastasis, osteoporosis, and the development of late complications of diabetes, e.g., vascular hyperpermeability, nephropathy, retinopathy and neuropathy.

As used herein, the term "subject" refers to a mammalian subject, preferably a human, suffering from or at risk of suffering from one or more of the above diseases or disease states.

Compounds that antagonize the interaction of RAGE with its physiological ligands are potentially useful in treating diseases or conditions that may respond to inhibition of the RAGE receptor. The invention provides a method of treatment comprising administering to a subject a compound of the invention or a pharma- ceutically acceptable salt thereof. In an embodiment of this aspect, the invention provides a method of inhibiting the interaction of RAGE with its physiological ligand. In another embodiment of this aspect, the invention provides a method of inhibiting RAGE signaling.

In another aspect, the present invention provides a method for treating a RAGE-mediated disease comprising administering a compound of the present invention or a pharma- ceutically acceptable salt thereof. In another aspect, the present invention provides a method for treating a RAGE-mediated disease comprising administering a therapeutically effective amount of a compound of the present invention or a pharma-ceutically acceptable salt thereof.

I. RAGE and Diabetic Complications As noted above, the compounds of the present invention may be useful in treating diabetic complications. Nonenzymatic glycoxidation of macromolecules, ultimately resulting in the formation of advanced glycation end products (AGEs), has been shown to be enhanced in renal failure, in the presence of hyperglycemia and other conditions associated with systemic or local oxidant stress, and at sites of inflammation (Dyer, D., et al., J. Clin. Invest., 91:2463-2469 (1993); Reddy, S., et al., Biochem., 34:10872-10878 (1995); Dyer, D. et al., J. Biol. Chem., 266:11654-11660 (1991); Degenhardt, T., et al., Cell Mol. Biol., 44:1139-1145 (1998)). Accumulation of AGEs in the vasculature can occur locally, as in the joint amyloid composed of AGE-β2-microglobulin found in patients with dialysis-associated amyloidosis (Miyata, T., et al., J. Clin. Invest., 92:1243-1252 (1993); Miyata, T., et al., J. Clin. Invest., 98:1088-1094 (1996)), or as exemplified by the vasculature and tissues of diabetic patients in general (Schmidt, A-M., et al., Nature Med., 1:1002-1004 (1995)). The progression of AGE accumulation over time in diabetic patients suggests that endogenous clearance mechanisms are unable to function effectively at the sites of AGE deposition. These accumulated AGEs have the ability to alter cellular properties by several mechanisms. RAGE is expressed at low levels in normal tissues and vasculature, but in environments where the receptor's ligand accumulates, RAGE has been shown to be upregulated (Li, J., et al., J. Biol. Chem., 272:16498-16506 (1997); Li, J., et al., J. Biol. Chem., 73:30870-30878 (1998); Tanaka, N., et al., J. Biol. Chem., 275:25781-25790 (2000)). RAGE expression is increased in endothelium, smooth muscle cells, and infiltrating mononuclear phagocytes in the diabetic vasculature. Studies in cell culture have also demonstrated that AGE-RAGE interactions induce changes in cellular properties important for vascular homeostasis.

II. RAGE and Cellular Dysfunction in Amyloidosis As also described above, the compounds of the present invention may be useful in the treatment of amyloidosis and/or Alzheimer's disease. RAGE appears to be a cell surface receptor that binds to β-sheet fibrillar material regardless of the composition of the subunits (amyloid-β peptide, Aβ, amylin, serum amyloid A, prion-derived peptides) (Yan, S.-D., et al., Nature, 382:685-691 (1996); Yan, S.-D., et al., Nat. Med., 6:643-651 (2000)). Amyloid deposition has been shown to result in enhanced expression of RAGE. For example, in the brains of Alzheimer's disease (AD) patients, RAGE expression is increased in neurons and glia (Yan, S.-D., et al., Nature 382:685-691 (1996)). The outcome of Aβ-RAGE interaction appears to be quite different in neurons and microglia. Microglia are activated as a result of Aβ-RAGE interaction, but early RAGE-mediated neuronal activation is replaced by late cytotoxicity, as reflected by increased motility and expression of cytokines. Further evidence for the role of RAGE in Aβ cellular interactions concerns the inhibition of Aβ-induced cerebral vasoconstriction and translocation of the peptide across the blood-brain barrier into the brain parenchyma when the receptor is blocked (Kumar, S., et al., Neurosci. Program, p141 (2000)). Inhibition of RAGE-amyloid interactions has been shown to decrease cellular RAGE and cell stress marker expression (as well as NF-kB activation) and reduce amyloid deposition (Yan, S-D., et al., Nat. Med., 6:643-651 (2000)), suggesting a role for RAGE-amyloid interactions in both the perturbation of cellular properties and amyloid accumulation in amyloid-rich environments (even at early stages).

Additionally, both cell assays and animal studies have shown that RAGE mediates transcytosis of circulating Aβ across the blood-brain barrier (BBB). This increased transcytosis of Aβ results in neuronal oxidant stress and a sustained decrease in cerebral blood flow. The action of RAGE can be inhibited by RAGE modulators, such as anti-RAGE antibodies or sRAGE (see, e.g., Mackic et al., J. Clin. Invest., 102:734-743 (1998); see also Kumar et al., Neurosci., Program, p141 (2000)). These findings were confirmed by further studies (see, e.g., U.S. Pat. No. 6,825,164, col. 17, line 48 to col. 18, line 43; Deane et al., Nature Medicine, 9:907-913 (2003)). Reduced cerebral perfusion can promote ischemic lesions that may act synergistically with Aβ to exacerbate dementia. Insufficient cerebral blood flow may also alter Aβ transport across the blood-brain barrier, thereby decreasing Aβ clearance and promoting Aβ accumulation in the brain (see Girouard and Iadecola, J. Appl. Physiol., 100, 328-335 (2006) at p. 332). Thus, increased cerebral blood flow promoted by RAGE antagonists may alleviate symptoms of Alzheimer's disease, delay its onset, or both. For example, RAGE antagonists may delay or slow the loss of cognitive ability, or may improve cognitive ability in subjects with Alzheimer's dementia, or both.

III. RAGE and Propagation of Immune/Inflammatory Responses As noted above, the compounds of the present invention can be useful in the treatment of inflammation. For example, S100/calgranulin has been shown to comprise a family of closely related calcium-binding polypeptides characterized by two EF-hand domains linked by a binding peptide (Schafer, B. et al., TIBS, 21:134-140 (1996); Zimmer, D., et al., Brain Res. Bull., 37:417-429 (1995); Rammes, A., et al., J. Biol. Chem., 272:9496-9502 (1997); Lugering, N., et al., Eur. J. Clin. Invest., 25:659-664 (1995)). Although S100/calgranulin lacks a signal peptide, it has long been known to gain access to the extracellular space, particularly at sites of chronic immune/inflammatory responses such as cystic fibrosis and rheumatoid arthritis. RAGE is the receptor for many members of the S100/calgranulin family and mediates proinflammatory effects on cells such as lymphocytes and mononuclear phagocytes. Studies on delayed-type hypersensitivity reactions, colitis in IL-10 null mice, collagen-induced arthritis, and experimental autoimmune encephalitis models also suggest that RAGE-ligand interactions (possibly with S100/calgranulin) have a proximal role in inflammatory cascades such as those involved in inflammatory diseases, including but not limited to rheumatoid arthritis and multiple sclerosis.

RAGE is also involved in inflammatory diseases of the skin, including, but not limited to, atopic dermatitis, eczema, and psoriasis. In particular, psoriasis is characterized by inflammatory and itchy lesions. Psoriasis may be accompanied by arthritic symptoms similar to those seen in rheumatoid arthritis. There is considerable evidence that psoriasis is a polygenic autoimmune disease. Psoriatic lesions are rich in cytokines, particularly IL-1 and IL-8, both of which are potent proinflammatory mediators. In particular, IL-8 is a chemotactic factor for neutrophils; neutrophils are also known to synthesize and secrete S100 proteins, which are one of the ligands for RAGE involved in the propagation of immune and inflammatory responses. Psoriasin (S100A7), a novel member of the S100 gene family, is a secreted protein isolated from psoriatic skin. Semprini et al. , (Hum. Genet. 2002 Oct, 111(4-5), 310-3) showed a link between genetic susceptibility to psoriasis and a distinct overexpression of S100 proteins in the skin. Therefore, modulators of RAGE are expected to regulate the immune response in psoriasis.

IV. RAGE and Amphoterin As noted above, the compounds of the present invention can be useful in the treatment of tumors and tumor metastases. For example, amphoterin is a high mobility group I non-histone chromosomal DNA binding protein that has been shown to interact with RAGE (Rauvala, H., et al., J. Biol. Chem., 262:16625-16635 (1987); Parkikinen, J., et al., J. Biol. Chem. 268:19726-19738 (1993)). Amphoterin has been shown to promote neurite outgrowth as well as to serve as a surface for assembly of protease complexes in the fibrinolytic system (also known to contribute to cell motility). Furthermore, local tumor growth inhibitory effects of blocking RAGE have been observed in a primary tumor model (C6 glioma), a Lewis lung metastasis model (Taguchi, A., et al., Nature 405:354-360 (2000)), and in spontaneously arising papillomas in mice expressing the v-Ha-ras transgene (Leder, A., et al., Proc. Natl. Acad. Sci., 87:9178-9182 (1990)).

V. RAGE and Respiratory Disease Airway inflammation is important in the pathogenesis of asthma. Such inflammation can lead to significant exacerbations and increases in the severity of asthma, as well as being a major factor in the reduction of asthma condition. Severe asthma exacerbations are characterized by a strong inflammatory response with heterogeneous mechanisms involving the accumulation and activation of neutrophils and eosinophils. Neutrophils are an important source of S100 proteins, the main ligands of RAGE, which are involved in immune responses and the propagation of inflammation. Therefore, modulators of RAGE are expected to have therapeutic value in the treatment of asthma.

Furthermore, the propagation phase of the immune response in the lung driven by S100-RAGE interactions is expected to result in the activation and/or recruitment of inflammatory cells such as neutrophils, which are a significant source of damaging proteases in chronic obstructive pulmonary diseases such as emphysema. Thus, RAGE modulators are expected to have potential in the treatment of chronic obstructive pulmonary diseases.

As used herein, the phrase "therapeutically effective amount" is intended to mean an amount of a drug or pharmaceutical agent that elicits a desired therapeutic response in a subject. In these methods, factors that may affect what constitutes a therapeutically effective amount include, but are not limited to, the size and weight of the subject, the biodegradability of the therapeutic agent, the activity of the therapeutic agent, the size of the area of action, and its bioavailability. The phrase encompasses an amount that results in improved treatment, cure, or amelioration of side effects, or a reduction in the rate of progression of a disease or disorder, as compared to a corresponding subject not receiving such amount.

As used herein, the term "treatment" refers to the full range of treatments for a given condition or disorder from which a subject suffers, for example, from alleviating or ameliorating one or more symptoms resulting from the disorder, to delaying the onset or progression of the disorder.

In one embodiment, the present invention provides a method for treating restenosis, comprising administering to a subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutical acceptable salt thereof. In one embodiment, the subject suffers from diabetes.

In one aspect, the invention provides a method for treating acute or chronic inflammation, comprising administering to a subject a therapeutically effective amount of a compound of the invention, or a pharma- ceutically acceptable salt thereof.
In one embodiment, the present invention provides a method for treating dementia associated with head trauma, comprising administering to a subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof. In one embodiment, the subject's cognitive ability is improved. In another embodiment, the subject's cognitive ability is maintained. In another embodiment, the rate of loss of cognitive ability in the subject is slowed.

In one aspect, the present invention provides a method for treating Alzheimer's disease, comprising administering to a subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof. With respect to Alzheimer's disease, the present invention is believed to be useful in altering the course of the underlying dementia process. Alzheimer's disease can be diagnosed within certain limits by NINCDS and DSM criteria, the Mini-Mental State Examination, and the Clinical Dementia Rating. One aspect of the present invention includes improving cognitive performance, comprising administering a compound of the present invention or a pharma- ceutically acceptable salt thereof. Cognitive performance can be assessed by the cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-cog), as known in the art, which scores cognitive function on a scale of 0 to 70, with higher scores indicating greater cognitive impairment. Thus, a decrease in score indicates cognitive improvement. One aspect of the present invention includes administering to a subject a compound of the present invention or a pharma- ceutically acceptable salt thereof to reduce the ADAS-cog score of a subject in need of such a reduction. Such a subject can be a human suffering from Alzheimer's dementia, mild to moderate Alzheimer's disease, or severe Alzheimer's disease.

In addition, the progression of Alzheimer's disease can also be assessed in humans through examination of four domains of functioning: global, cognition, behavior, and activities of daily living. Such assessments can be performed using the Clinical Impression of Change in Dementia (CIBIC or CIBIC Plus). One aspect of the invention includes improving a subject's functioning, comprising administering a compound of the invention or a pharmacologic acceptable salt thereof. In one embodiment, the subject's functioning is one or more of global, cognition, behavior, and activities of daily living.

In one aspect, the present invention provides a method for improving wound healing in a diabetic subject, comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof to improve the rate of wound healing in the subject as compared to an untreated wound.

In one embodiment, the present invention provides a method for treating inflammation associated with transplantation of an organ, tissue, or multiple cells from a first site to a second site in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof to reduce inflammation in the subject. In one embodiment, the first and second sites are in different subjects. In another embodiment, the first and second sites are in the same subject. In another embodiment, the transplanted organ, cell, or tissue comprises a cell or tissue of the pancreas, skin, liver, kidney, heart, bone marrow, blood, bone, muscle, artery, vein, cartilage, thyroid, nervous system, or stem cell.

Combination Therapy In another embodiment, at least one compound of the present invention, or a pharma- ceutically acceptable salt thereof, is utilized alone or in combination with one or more known therapeutic agents, including azeliragon, or a pharma- ceutically acceptable salt thereof.

In a further aspect of the invention, the compounds of the invention can be used in adjuvant therapy or in combination therapeutic treatment with other known therapeutic agents.
The following is a non-exhaustive list of adjuvants and additional therapeutic agents that can be utilized in combination with the compounds of the invention:

Pharmacological classification of anticancer drugs:
1. Alkylating agents: cyclophosphamide, nitrosoureas, carboplatin, cisplatin, procarbazine 2. Antibiotics: bleomycin, daunorubicin, doxorubicin 3. Antimetabolites: methotrexate, cytarabine, fluorouracil 4. Plant alkaloids: vinblastine, vincristine, etoposide, paclitaxel 5. Hormones: tamoxifen, octreotide acetate, finasteride, flutamide 6. Biological response modifiers: interferons, interleukins, antitumor antibodies

Pharmacological classification of treatment for rheumatoid arthritis (inflammation) 1. Analgesics: Aspirin 2. NSAIDs (Non-steroidal anti-inflammatory drugs): Ibuprofen, Naproxen, Diclofenac 3. DMARDs (Disease-modifying antirheumatic drugs): Methotrexate, Gold preparations, Hydroxychloroquine, Sulfasalazine 4. Biological response modifiers, DMARDs: Etanercept, Infliximab, Glucocorticoids

Pharmacological Classification of Diabetes Treatment 1. Sulfonylureas: Tolbutamide, Tolazamide, Glyburide, Glipizide 2. Biguanides: Metformin 3. Various Oral Agents: Acarbose, Troglitazone 4. Insulin

Pharmacological Classification of Treatments for Alzheimer's Disease 1. Cholinesterase Inhibitors: Tacrine, Donepezil 2. Antipsychotics: Haloperidol, Thioridazine 3. Antidepressants: Desipramine, Fluoxetine, Trazodone, Paroxetine 4. Anticonvulsants: Carbamazepine, Valproic Acid

In a further aspect, the present invention provides a method for treating a RAGE-mediated disease, the method comprising administering to a subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof in combination with a therapeutic agent selected from the group consisting of alkylating agents, antimetabolites, plant alkaloids, antibiotics, hormones, biological response modifiers, analgesics, NSAIDs, DMARDs, glucocorticoids, sulfonylureas, biguanides, insulin, cholinesterase inhibitors, antipsychotics, antidepressants, and anticonvulsants.

In a further aspect, the present invention provides a pharmaceutical composition of the present invention as described above, further comprising one or more therapeutic agents selected from the group consisting of alkylating agents, antimetabolites, plant alkaloids, antibiotics, hormones, biological response modifiers, analgesics, NSAIDs, DMARDs, glucocorticoids, sulfonylureas, biguanides, insulin, cholinesterase inhibitors, antipsychotics, antidepressants, and anticonvulsants.

Such other therapeutic agents may be administered by the same route as the compound of the invention or a pharma- ceutically acceptable salt thereof or by a different route. When the compound of the invention or a pharma- ceutically acceptable salt thereof is used in combination with other therapeutic agents, the composition may contain the compound of the invention or a pharma- ceutical acceptable salt thereof in combination with the other therapeutic agent(s). Alternatively, when separate dosage formulations are used, the compound of the invention or a pharma- ceutical acceptable salt thereof and the one or more additional therapeutic agents may be administered at substantially the same time (e.g., simultaneously) or at separate staggered times (e.g., sequentially).

In any of the above embodiments of the method of treatment, the method may further comprise administering a compound of the present invention or a pharma- ceutically acceptable salt thereof and Compound I or a pharma- ceutically acceptable salt thereof. When administered together in a single dosage form or in separate dosage forms, the amount of Compound I or a pharma- ceutically acceptable salt thereof administered per dose may be 0.1 mg to 20 mg, and the amount of the compound of the present invention or a pharma- ceutically acceptable salt thereof may be 0.1 mg to 1000 mg. In other embodiments, the amount of Compound I or a pharma- ceutically acceptable salt thereof administered per dose may be 0.1 mg to 20 mg, and the amount of the compound of the present invention or a pharma- ceutical acceptable salt thereof may be 0.0001% to 5%, or 0.001% to 5%, by weight, relative to the amount of Compound I or a pharma- ceutical acceptable salt thereof, or relative to the total weight of the pharmaceutical composition.

Pharmaceutical Formulations and Dosage Forms The present invention further provides pharmaceutical compositions comprising the compounds of the present invention or pharma- ceutically acceptable salts thereof and a pharma- ceutical acceptable carrier. As used herein, the term "pharmaceutical composition" is used to denote a composition that can be administered to a mammalian host, for example, orally, topically, parenterally, by inhalation spray, or rectally, in a unit dosage formulation containing conventional non-toxic carriers, diluents, adjuvants, vehicles, and the like. As used herein, the term "parenterally" includes subcutaneous injection, intravenous, intramuscular, intracisternal injection, or by infusion techniques.

Pharmaceutical compositions containing the compounds of the invention can be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use can be prepared according to any known method, and such compositions can contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives, in order to provide medicamentously elegant and palatable preparations. Tablets can contain the active ingredient in admixture with non-toxic pharma-ceutically acceptable excipients suitable for the manufacture of tablets. These excipients can be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricants, such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be employed. They may also be coated by techniques to form osmotic therapeutic tablets for control release.

Formulations for oral use can also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions may contain the active compound in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic; dispersing or wetting agents may be naturally occurring phosphatides, for example, lecithin, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, for example, heptadecaethyl-enoxycetanol, or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitols, for example, polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitol anhydrides, for example, polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, for example, sucrose or saccharin.

Oily suspensions can be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions can contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavouring agents can be added to provide a palatable oral preparation. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active compound in admixture with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, such as gum arabic or gum tragacanth, naturally occurring phosphatides, such as soybean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.

Syrups or elixirs can be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol or sucrose. Such formulations can also contain demulcents, preservatives, flavoring and coloring agents. The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oily suspension. The suspensions can be formulated according to known methods using suitable dispersing or wetting agents and suspending agents, as described above. The sterile injectable preparations can also be as a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conveniently employed as a solvent or suspending medium. For this purpose, any non-irritating fixed oil can be employed, using synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid, find use in the preparation of injectable preparations.

The compositions may also be in the form of suppositories for rectal administration of the compounds of the invention. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at rectal temperature and will therefore melt in the rectum to release the drug. Such materials include, for example, cocoa butter and polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, lotions, eye ointments and ear drops, impregnated bandages, aerosols, and the like, containing the compounds of the invention are contemplated. These topical formulations can contain suitable conventional additives, such as preservatives, solvents to aid drug penetration, and emollients in ointments and creams. The formulations can also contain compatible conventional carriers, such as cream or ointment bases, and ethanol or oleyl alcohol for lotions. Such carriers can be present from about 0.1% to about 99% of the formulation. More usually, they will form up to about 80% of the formulation. For purposes of this application, topical applications include mouthwashes and gargles.

The compounds of the invention can also be coupled to soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymers, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethylene oxide polylysine substituted with palmitoyl residues. Additionally, the compounds of the invention can be coupled to classes of biodegradable polymers useful for achieving controlled release of drugs, such as polylactides, polyepsiloncaprolactones, polyhydroxybutyrates, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and crosslinked or amphiphilic block copolymers of hydrogels.

For administration by inhalation, the compounds according to the invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer using a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, tetrafluoroethane, heptafluoropropane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, e.g., gelatin, for use in an inhaler or insufflator can be formulated containing a powder mix of the compound of the invention and a suitable powder base such as lactose or starch.

Generally speaking, the compounds of the invention or pharma- ceutically acceptable salts thereof can be administered at dosage levels of about 0.003-500 mg/kg body weight of the subject being treated. In one embodiment, the compounds of the invention or pharma- ceutically acceptable salts thereof can be administered at a dosage range of about 0.003-200 mg/kg body weight per day, every other day, or once a week. In one embodiment, the compounds of the invention or pharma- ceutically acceptable salts thereof can be administered at a dosage range of about 0.1 mg-100 mg per day, every other day, or once a week.

The amount of active ingredient that can be combined with the carrier material to produce a single dosage can vary depending on the host treated and the particular mode of administration. For example, a formulation intended for oral administration to humans can contain 1 mg to 2 grams of the compound of the present invention or a pharma- ceutically acceptable salt thereof, together with an appropriate and convenient amount of carrier material, which can vary from about 5 to 95 percent of the total composition. Dosage forms intended for topical administration to the skin can be prepared with a ratio of compound to topical excipient of 0.1% to 99%. Dosage forms intended for inhalation administration are 0.01 to 200 mg of compound in a carrier suitable for delivering an inhaled dose of the compound. A unit dosage form of the compound delivered systemically can generally contain about 1 mg to about 500 mg of active ingredient. This dosage can be individualized by the clinician based on the specific clinical condition of the subject being treated. It will therefore be understood that the specific dosage level for any particular subject will depend on a variety of factors, including the activity of the specific compound employed, age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, size of the spectrum of action, and the severity of the particular disease being treated.

In addition to being a metabolite of Compound I, M3 has also been found to be a degradation product of Compound I and/or an impurity resulting from certain synthetic methods of Compound I. Thus, in another aspect, the present invention provides a pharmaceutical composition comprising M3 or a pharma- ceutically acceptable salt thereof, Compound I or a pharma-ceutically acceptable salt thereof, and a pharma-ceutically acceptable carrier. In another aspect, the present invention provides a pharmaceutical composition comprising M3 or a pharma-ceutically acceptable salt thereof, and Compound I or a pharma-ceutically acceptable salt thereof, wherein M3 or a pharma-ceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of 0.0001% to 5% by weight of the pharmaceutical composition. In another aspect, M3 or a pharma-ceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of 0.01% to 0.5% by weight of the pharmaceutical composition. In other embodiments, the pharmaceutical composition comprises M3 or a pharma- ceutically acceptable salt thereof and 0.1 mg to 100 mg, or 0.1 mg to 50 mg, or 1 mg to 30 mg, or 1 mg to 10 mg of Compound I or a pharma- ceutically acceptable salt thereof, and M3 or a pharma- ceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of 0.0001% to 5%, or 0.001% to 0.01%, or 0.01% to 1% by weight relative to the amount of Compound I or a pharma- ceutical acceptable salt thereof in the pharmaceutical composition or relative to the total weight of the pharmaceutical composition.

Labeled Compounds and Assay Methods Another aspect of the invention pertains to labeled compounds (radiolabeled, fluorescently labeled, etc.) of the invention that are useful in assays, as well as imaging techniques, both in vitro and in vivo.

The present invention further includes isotopically labeled or isotopically labeled compounds of the present invention. An "isotopically" or "radiolabeled" compound is a compound of the present invention in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that can be incorporated into the compounds of the present invention include, but are not limited to, ² H (also written as D for deuterium), ³ H (also written as T for tritium), and ¹⁴ C. The radionuclides incorporated into the present radiolabeled compounds depend on the specific application of the radiolabeled compounds.

The labeled compounds of the invention can be used in screening assays or toxicological or pharmacokinetic studies of Compound I to identify/assess the absorption, distribution, metabolism, and excretion profiles of Compound I and/or its metabolites. For example, isotopically enriched forms of various metabolites can be prepared by substituting d ₉ -valeryl chloride (CD ₃ CD ₂ CD ₂ CD ₂ COCl) for valeryl chloride during synthesis of the metabolites, such that the butyl groups of M1-M3 and M5-M7 become isotopically enriched with deuterium.
The present specification includes the following aspects of the invention.
[1] A compound selected from the group consisting of:
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine;
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol;
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol; and
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol;
or a pharma- ceutically acceptable salt thereof.
[2] The compound according to [1], wherein the compound is 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid or a pharma- ceutically acceptable salt thereof.
[3] The compound according to [1], wherein the compound is [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine or a pharma- ceutically acceptable salt thereof.
[4] The compound according to [1], wherein the compound is 2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol or a pharma- ceutically acceptable salt thereof.
[5] The compound according to [1], wherein the compound is 5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol or a pharma- ceutically acceptable salt thereof.
[6] The compound according to [1], wherein the compound is 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol or a pharma- ceutically acceptable salt thereof.
[7] A compound selected from the group consisting of:
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine;
4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol;
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol;
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol; and
3-(4-{2-{butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol;
or a pharma- ceutically acceptable salt thereof; and
at least one pharma- ceutically acceptable carrier;
13. A pharmaceutical composition comprising:
[8] 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol or a pharma- ceutically acceptable salt thereof;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine or a pharma- ceutically acceptable salt thereof; and
A pharma- ceutically acceptable carrier;
13. A pharmaceutical composition comprising:
[9] 0.1 mg to 100 mg of [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine or a pharma- ceutically acceptable salt thereof, and
0.0001% to 5% by weight of 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol or a pharma- ceutically acceptable salt thereof
The pharmaceutical composition according to [8],
[10] A method for treating skin inflammation, such as psoriasis, rheumatoid arthritis, atopic dermatitis, and pulmonary inflammation, such as acute or chronic inflammation including asthma and chronic obstructive pulmonary disease, diabetes, diabetes-related complications, renal failure, hyperlipidemic atherosclerosis associated with diabetes, neuronal toxicity, restenosis, Down's syndrome, dementia associated with head trauma, amyotrophic lateral sclerosis, multiple sclerosis, amyloidosis, autoimmune diseases, such as inflammation associated with autoimmunity or organ, tissue or cell transplantation, wound healing disorders, periodontal disease, neuropathy, neuronal degeneration, vascular permeability, nephropathy, atherosclerosis, retinopathy, Alzheimer's disease, erectile dysfunction, tumor invasion and/or metastasis, osteoporosis, and the development of late complications of diabetes, such as vascular hyperpermeability, nephropathy, retinopathy and neuropathy,
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine;
4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol;
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol;
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol; and
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol;
or a pharma- ceutically acceptable salt thereof;
A method comprising administering to a subject a compound selected from the group consisting of:
[11] 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine;
4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol;
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol;
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol; and
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol;
or a pharma- ceutically acceptable salt thereof;
20. Use of a compound selected from the group consisting of: in the preparation of a medicament.
[12] 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid;
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine;
4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol;
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol;
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol; and
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol;
or a pharma- ceutically acceptable salt thereof;
A compound selected from the group consisting of
Inflammation of the skin, e.g. psoriasis, rheumatoid arthritis, atopic dermatitis, and inflammation of the lungs, e.g. acute or chronic inflammation including asthma and chronic obstructive pulmonary disease, diabetes, diabetes-related complications, renal failure, hyperlipidemic atherosclerosis associated with diabetes, neuronal toxicity, restenosis, Down's syndrome, dementia associated with head trauma, amyotrophic lateral sclerosis, multiple sclerosis, amyloidosis, autoimmune diseases, e.g. inflammation associated with autoimmunity or organ, tissue or cell transplantation, wound healing disorders, periodontal disease, neuropathy, neuronal degeneration, vascular permeability, nephropathy, atherosclerosis, retinopathy, Alzheimer's disease, erectile dysfunction, tumor invasion and/or metastasis, osteoporosis, and the development of late complications of diabetes, e.g. vascular hyperpermeability, nephropathy, retinopathy and neuropathy.
In the preparation of a medicament for treating

Metabolite 1 (M1): 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid

To 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol (5.5 g) (see US Pat. Nos. 7,361,678 and 8,580,833) in t-butyl acrylate (50 mL) was added DBU (1 eq.) and the mixture was heated at 100° C. for 3 h. The reaction mixture was concentrated and water (100 mL) and a 1:1 ethyl acetate-hexane mixture (100 mL) were added. The mixture was stirred for 30 min and filtered. The organic layer was isolated and concentrated. The residue was purified by column chromatography on silica gel using 1-100% ethyl acetate in hexane as eluent. The desired fractions were concentrated and the residue was co-evaporated with n-heptane to give tert-butyl 3-[4-[2-butyl-1-[4-(4-chlorophenoxy)phenyl]imidazol-4-yl]phenoxy]propanoate (1.4 g). The ester was dissolved in 4N HCl in dioxane. Water (5 mL) was added and the mixture was stirred for 30 min. The resulting solution was concentrated and dried to give 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid as a solid.
^1H NMR (DMSO- _d6 , TMS): δ 0.82 (t, 3H), 1.25 (m, 2H), 1.59 (m, 2H), 2.75 (t, 2H), 2.92 (m, 2H), 4.22 (t, 2H), 7.12-7.20 (m, 4H), 7.29 (d, 2H), 7.55 (d, 2H), 7.75 (d, 2H), 7.92 (d, 2H), 8.21 (s, 1H), ppm.
Metabolite 2 (M2): [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethylamine

[3-(4-{2-{butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine (18 g) (see US Pat. No. 7,361,678) was dissolved in anhydrous dichloromethane (150 mL) in a 3-neck round bottom flask under nitrogen. The contents were cooled to -78°C and a solution of (1-chloroethyl)-chloroformate (100 g) in dichloromethane (150 mL) was added dropwise over 0.5 hours. The contents were allowed to warm to room temperature and stirred at that temperature for 16 hours. All volatiles were evaporated under reduced pressure on a rotavapor. Water (150 mL) and ethyl acetate were added and the pH was adjusted to about 12 with 5N sodium hydroxide with stirring. The layers were separated and the organic layer was dried over sodium sulfate and concentrated. The residue was purified by column chromatography on silica gel using 5-15% methanol in dichloromethane as eluent. The desired fractions were concentrated and the residue was co-evaporated with n-heptane to give [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethylamine as a solid (7.8 g).
^1H NMR ( _CDCl3 , TMS): δ 0.82 (t, 3H), 1.15 (t, 3H), 1.35 (m, 3H), 1.65 (m, 2H), 1.95 (m, 2H), 2.65 (m, 4H), 2.8 (t, 2H), 4.05 (t, 2H), 6.9 (d, 2H), 7.05 (d, 2H), 7.1 (d, 2H), 7.17 (s, 1H), 7.25 (d, 2H), 7.35 (d, 2H), 7.75 (d, 2H) ppm.
Metabolite 3 (M3): 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol

See Intermediate A1 of US Pat. No. 8,580,833.
Metabolite 5 (M5): 2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol

N-[4-[4-chloro-2-[(4-methoxyphenyl)methoxy]phenoxy]phenyl]-N-[2-(4-hydroxyphenyl)-2-oxo-ethyl]pentanamide (1.3 g) was prepared from 4-acetoxyacetophenone, 4-[4-chloro-2-[(4-methoxyphenyl)methoxy]phenoxy]aniline and valeroyl chloride using previously described procedures (see U.S. Pat. Nos. 7,361,678 and 8,580,833) and dichloromethane. It was converted to N-[4-[4-chloro-2-[(4-methoxyphenyl)methoxy]phenoxy]phenyl-N-[2-[4-[3-(diethylamino)propoxy]phenyl]-2-oxo-ethyl]pentanamide (0.8 g) by treatment with 3-(diethylamino)propan-1-ol, triphenylphosphine and DIAD in dichloromethane, followed by column purification on silica using 0-15% methanol in dichloromethane as eluent.

N-[4-[4-chloro-2-[(4-methoxyphenyl)methoxy]phenoxy]phenyl]-N-[2-[4-[3-(diethylamino)propoxy]phenyl]-2-oxo-ethyl]pentanamide (0.8 g) was converted to 2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol (1.2 g) by treatment with ammonium acetate in acetic acid at 100-110° C. for 48 hours using the procedure previously described (see U.S. Pat. No. 7,361,678).
^1H NMR ( _CDCl3 , TMS): δ 0.65 (t, 3H), 1.25 (t, 6H), 1.25 (m, 2H), 1.65 (m, 2H), 1.95 (s, 1H), 2.05 (m, 2H), 2.52 (m, 2H), 2.75-2.9 (m, 6H), 3.92 (t, 2H), 6.85-6.92 (m, 4H), 7.02-7.12 (m, 4H), 7.2 (d, 2H), 7.64 (d, 2H) ppm.
Metabolite 6 (M6): 5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol

N-[4-[4-chloro-3-[(4-methoxyphenyl)methoxy]phenoxy]phenyl]-N-[2-(4-hydroxyphenyl)-2-oxo-ethyl]pentanamide (5.36 g) was prepared from 4-acetoxyacetophenone, 4-[4-chloro-3-[(4-methoxyphenyl)methoxy]phenoxy]aniline and valeroyl chloride using previously described procedures (see U.S. Pat. Nos. 7,361,678 and 8,580,833) and dichloromethane. It was converted to N-[4-[4-chloro-3-[(4-methoxyphenyl)methoxy]phenoxy]phenyl-N-[2-[4-[3-(diethylamino)propoxy]phenyl]-2-oxo-ethyl]pentanamide (5.1 g) by treatment with 3-(diethylamino)propan-1-ol, triphenylphosphine and DIAD in dichloromethane, followed by column purification on silica using 0-15% methanol in dichloromethane as eluent.

N-[4-[4-chloro-3-[(4-methoxyphenyl)methoxy]phenoxy]phenyl-N-[2-[4-[3-(diethylamino)propoxy]phenyl]-2-oxo-ethyl]pentanamide (5.1 g) was converted to 5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol (1.2 g) by treatment with ammonium acetate in acetic acid at 100-110° C. for 48 hours using the procedure previously described (see U.S. Pat. No. 7,361,678).
^1H NMR ( _CDCl3 , TMS): δ 0.65 (t, 3H), 1.2 (t, 6H), 1.25 (t, 2H), 1.65 (m, 2H), 2.0 (m, 2H), 2.6-2.75 (m, 8H), 3.2 (br, 1H), 4.05 (t, 2H), 6.56 (m, 1H), 6.7 (m, 1H), 6.88 (d, 2H), 7.04 (m, 3H), 7.25 (d, 2H), 7.3 (d, 1H), 7.66 (d, 2H) ppm.
Metabolite 7 (M7): 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol

A mixture of 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol (3 g) (see U.S. Pat. Nos. 7,361,678 and 8,580,833), 3-chloropropanol and potassium carbonate in acetone (50 mL) was heated to reflux for 3 days. The reaction mixture was filtered, concentrated and the residue was purified by column chromatography on silica gel using 1-100% ethyl acetate in hexane as eluent. The desired fractions were concentrated and the residue was recrystallized from ethyl acetate and hexane to give 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol (2.7 g) as a white solid.
^1H NMR ( _CDCl3 , TMS): δ 0.85 (t, 3H), 1.32 (m, 2H), 1.65 (m, 2H), 1.8 (br, 1H), 2.05 (m, 2H), 2.65 (m, 2H), 3.78 (m, 2H), 4.25 (t, 2H), 6.92 (d, 2H), 7.02 (d, 2H), 7.08 (d, 2H), 7.03 (s, 1H), 7.29 (d, 2H), 7.35 (d, 2H), 7.71 (d, 2H) ppm.

Metabolism Study of Compound I Evaluation of the metabolism of Compound I was performed using human plasma and bile samples collected from subjects administered multiple doses of Compound I. This was a multiple-dose study evaluating eight healthy subjects who received Compound I orally for 14 days. Subjects received a loading dose of 15 mg once daily for 6 days, followed by a daily maintenance dose of 5 mg for 8 days. Plasma samples were collected before dosing on day 1. Plasma was also collected before dosing on days 11, 12, and 13. In addition, plasma was collected at 0, 1, 2, 4, 6, 8, and 10 hours after dosing on day 14, and at 24 and 28 hours after dosing on day 15. Bile samples were collected on day 15 (28 hours after dosing) by continuous suction through an oral enteral tube fluoroscopically placed near the bile duct for up to 4 hours. Bile was collected in the following fractions: 0-0.5 hours after the start of bile collection (approximately 24-24.5 hours after administration of Compound I), 0.5-1.0 hours, 1.0-2.0 hours, 2.0-3.0 hours, and 3.0-4.0 hours after the start of bile collection.

One assay method for determining the concentration of Compound I and certain metabolites in human plasma is as follows: The concentration of Compound I and certain metabolites in human plasma can be measured using an assay based on protein precipitation followed by high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS), with a lower limit of quantification of 0.2 ng/mL or 0.4 ng/mL for each analyte. An internal standard solution (0.5 mg/mL in methanol-DMSO, 90:10 v/v) can be prepared using deuterated (d ₉ ) Compound I, M1, M2, M3, M5, M6 or M7 internal standard compounds. An aliquot of 50 μL of K ₂ EDTA plasma is added to a 2 mL 96 DeepWell® plate containing 450 μL of blank solvent (acetonitrile containing 0.2% formic acid, v/v) and 450 μL of internal standard. The plate is vortexed at 1000 rpm for at least 5 minutes, followed by centrifugation at approximately 4000 rpm for at least 5 minutes at room temperature. An aliquot of 100 μL of the supernatant is transferred to either a 1 mL or 500 μL DeepWell® plate pre-filled with 100 μL of reconstitution solution (water-formic acid, 100:0.2 v/v). The plate is vortexed at approximately 1000 rpm for at least 5 minutes and then loaded onto an HPLC-MS/MS system for analysis. The HPLC conditions are as follows: column, XBridge (Waters) C18, 3.5 μM, 2.1×50 mm; column temperature, 45° C.; mobile phase composition, mobile phase A: 0.1% formic acid and 0.05% trifluoroacetic acid (vol/vol/vol), mobile phase B: acetonitrile containing 0.1% formic acid and 0.05% trifluoroacetic acid (vol/vol/vol); % of mobile phase A relative to % of mobile phase B varies from 65%/35% to 5%/95% over a measurement time of 12 minutes; flow rate, 1 mL/min. Quality control samples of plasma are analyzed at low, medium, and high concentrations in each analytical batch. Bile samples can be assayed using a similar method.

Microscale Thermophoresis The binding affinity of Compound I and the compounds of the present invention to human sRAGE (soluble extracellular domain of the human receptor for advanced glycation end products) was measured using microscale thermophoresis.

The binding affinity of compound I and metabolites M1, M2, M3, M5, M6 and M7 to recombinant human sRAGE was determined using a Microscale Thermophoresis System (Monolith NT.115 Pico, NanoTemper, Inc.). sRAGE was labeled with NT-647-NHS fluorescent dye. Approximately 5 nM of sRAGE was then added to compounds at concentrations ranging from 600 nM to 20 pM and incubated for 5 min at 25° C. in a buffer containing 25 mM Hepes (pH 7.5), 5 mM CaCl ₂ , 5 mM MgCl ₂ , 50 mM NaCl, 0.05% Tween 20 and 1% DMSO prior to microscale thermophoresis.

Raw thermophoretic data and average Kd were generated using Microscale Thermophoresis System analysis software following the manufacturer's protocol.

The average Kd between recombinant human sRAGE and compounds I, M1-M3, M5, M6 or M7 is shown in the table below.

S100B and Amyloid Beta ELISA Assays The following assays can be used to identify compounds that may be useful as inhibitors of the binding of physiological RAGE ligands, such as S100b and β-amyloid, to RAGE.

S100b or β-amyloid (500 ng/100 μL/well) in 100 mM sodium bicarbonate/sodium carbonate buffer (pH 9.8) is placed into the wells of a NUNC Maxisorp flat-bottom 96-well microtiter plate. The plate is incubated overnight at 4° C. The wells are aspirated and treated with 50 mM imidazole-buffered saline (pH 7.2) (with 5 mM CaCl ₂ /MgCl ₂ ) containing 1% bovine serum albumin (BSA) (300 μL/well) for 1 hour at room temperature. The wells are aspirated.

Test compounds are dissolved in nanopure water (concentration: 10-100 μM). DMSO can be used as a co-solvent. 25 μL of test compound solution in 4% DMSO is added to each well together with 75 μL of sRAGE (6 nM FAC) and the samples are incubated for 1 h at 37°C. The wells are washed several times with 155 mM NaCl buffered saline, pH 7.2, soaking for a few seconds between each wash.

Non-radioactive detection is performed by adding 10 μL of biotinylated goat F(ab') ₂ anti-mouse IgG (8.0× ^10-4 mg/mL, FAC), 5 μL Alk-phos-streptavidin (3× ^10-3 mg/mL FAC), and 0.42 μL per 5 mL of monoclonal antibody to sRAGE (FAC 6.0× ^10-3 mg/mL) to 5 mL of 50 mM imidazole-buffered saline (pH 7.2) containing 0.2% bovine serum albumin and 5 mM _CaCl2 . The mixture is incubated at room temperature for 30 minutes.

100 μL of complex is added to each well and incubation is allowed to proceed for 1 hour at room temperature. Wells are washed several times with wash buffer, soaking for a few seconds between each wash. 100 μL of 1 mg/mL (pNPP) in 1 M diethanolamine (pH adjusted to 9.8 with HCl) is added. Color is allowed to develop for 30 minutes to 1 hour at room temperature, protected from light. The reaction is quenched with 10 μL of stop solution (0.5-1.0 N NaOH in 50% ethanol) and absorbance is measured spectrophotometrically at 405 nm using a microplate reader.

Metabolite M2 was tested according to the assay method described above employing S100b or β-amyloid as the RAGE ligand and was found to have the following IC50s. The IC50 (μM) in the ELISA assay represents the concentration of compound at which 50% signal was inhibited.

Claims (10)

A compound selected from the group consisting of :
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid ,
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine ,
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol ,
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol , and 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol ,
or a pharma- ceutically acceptable salt thereof. The compound according to claim 1, wherein the compound is 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid or a pharma- ceutically acceptable salt thereof. The compound according to claim 1, wherein the compound is [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine or a pharma- ceutically acceptable salt thereof. The compound according to claim 1, wherein the compound is 2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol or a pharma- ceutically acceptable salt thereof. The compound according to claim 1, wherein the compound is 5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol or a pharma- ceutically acceptable salt thereof. The compound according to claim 1, wherein the compound is 3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol or a pharma- ceutically acceptable salt thereof. A compound selected from the group consisting of :
3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propionic acid ,
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-benzyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-ethyl-amine ,
4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol ,
2-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-5-chloro-phenol ,
5-[4-[2-butyl-4-[4-[3-(diethylamino)propoxy]phenyl]imidazol-1-yl]phenoxy]-2-chloro-phenol , and 3-(4-{2-{butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propan-1-ol ,
or a pharma- ceutically acceptable salt thereof , and at least one pharma- ceutically acceptable carrier ,
13. A pharmaceutical composition comprising: 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol or a pharma- ceutically acceptable salt thereof ,
[3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine or a pharma- ceutically acceptable salt thereof , and a pharma-ceutically acceptable carrier ,
13. A pharmaceutical composition comprising: 9. The pharmaceutical composition of claim 8, comprising 0.1 mg to 100 mg of [3-(4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenoxy)-propyl]-diethyl-amine or a pharma- ceutically acceptable salt thereof, and 0.0001% to 5% by weight of 4-{2-butyl-1-[4-(4-chloro-phenoxy)-phenyl]-1H-imidazol-4-yl}-phenol or a pharma- ceutically acceptable salt thereof. 10. The pharmaceutical composition according to any one of claims 7 to 9, for use in a method for treating skin inflammation , psoriasis , rheumatoid arthritis, atopic dermatitis , pulmonary inflammation , asthma , chronic obstructive pulmonary disease, acute or chronic inflammation, diabetes, diabetes-related complications, renal failure, hyperlipidemic atherosclerosis associated with diabetes, neuronal toxicity, restenosis, Down's syndrome, dementia associated with extracranial involvement, amyotrophic lateral sclerosis, multiple sclerosis, amyloidosis, autoimmune disease , inflammation associated with autoimmunity or organ, tissue or cell transplantation, wound healing disorders, periodontal disease, neuropathy, neuronal degeneration, vascular permeability, nephropathy, atherosclerosis, retinopathy, Alzheimer's disease, erectile dysfunction, tumor invasion and/or metastasis, osteoporosis , vascular permeability associated with diabetic late complications , nephropathy associated with diabetic late complications , retinopathy associated with diabetic late complications, or neuropathy associated with diabetic late complications .